M. tuberculosis strain-dependent interactions with host cells

结核分枝杆菌与宿主细胞的菌株依赖性相互作用

• 批准号: 10459539
• 负责人: JEFFERY S COX
• 金额: $ 45.63万
• 依托单位: UNIVERSITY OF WASHINGTON
• 依托单位国家: 美国
• 财政年份: 2021
• 资助国家: 美国
• 起止时间: 2021-08-01 至 2026-05-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10459539
• 关键词: Alleles; Animal Model; Bacterial Genes; Bacterial Proteins; Bioinformatics; Biological Assay; CBL gene; CRISPR/Cas technology; Candidate Disease Gene; Cell Wall; Cells; Clinical; Complement; Complex; Cues; Data; Data Set; Defect; Diagnostic; Disease; Disease Outcome; Distal; ELF3 gene; Etiology; Family; Genes; Genetic; Genetic Variation; Genotype; Goals; Growth; Host resistance; Human; Immune; Immune response; Individual; Infection; Inflammatory Response; Innate Immune Response; Interferon Type I; Interferons; Laboratories; M. tuberculosis genome; Mediating; Messenger RNA; Modification; Mutation; Mycobacterium tuberculosis; Network-based; Organism; Outcome; Pathogenesis; Pathogenicity; Pathway interactions; Phagosomes; Phenotype; Population; Post-Translational Protein Processing; Predisposition; Prevalence; Proteins; Proteomics; Pulmonary Tuberculosis; Resistance; Scanning; Site; System; Technology; Testing; Tuberculosis; Ubiquitin; Variant; Vietnam; Virulence; Work; bacterial genetics; base; causal variant; cohort; genetic testing; genome wide association study; high resolution imaging; human pathogen; in vivo; innate immune pathways; insight; macrophage; mouse model; mutant; pathogen; programs; recruit; repaired; response; transcriptomics; transmission process; ubiquitin ligase

Tuberculosis (TB) is a multifaceted disease that has extensive variation in clinical manifestations despite being the product of infection with a single pathogen, Mycobacterium tuberculosis. The extensive genetic diversity in human and Mycobacterium tuberculosis genomes responsible for differences in clinical outcomes represent modifications of host-pathogen interactions that are key to pathogenesis. Understanding the basis for these heterogenous responses will uncover new mechanisms of virulence and resistance and will impact treatment and diagnostics. Unfortunately, the challenges of studying the mechanisms of differential outcomes of infection in humans not only includes identification of correlations between host/pathogen genotypes with phenotypes in human populations, but also the subsequent identification of the mechanisms that are causal for disease which require studying them in the laboratory without the pathogen’s natural host organism. This Program takes advantage of unique, ongoing genome-wide association studies (GWAS) that have identified both human and pathogen variants that are associated with heterogenous clinical responses in two different human populations. To determine the mechanisms underlying these variations, we will employ a powerful set of experimental assays, including new proteomics-based scanning platform to probe host responses during experimental macrophage infection that is orthogonal to traditional mRNA profiling, in order to broadly search for changes in host innate immune pathways that correlate with disease outcomes associated with these clinical strains. Based on our preliminary data, we hypothesize that many of these interactions occur early during infection and are mediated by proteins secreted by M. tuberculosis. In this proposal, we focus primarily on correlations between two unique sets of clinical bacterial variants, strains that are associated greater transmission of pulmonary TB disease and strains that are more prone to dissemination to distal sites in the body. An unexpected theme from both of these sets of strains is the prevalence of TB proteins secreted by the ESX systems expressed in M. tuberculosis. Both the ESX-1 and ESX-5 secretion systems of M. tuberculosis are key virulence determinants required for intracellular growth and for eliciting distinct innate immune responses during macrophage infection. A central hypothesis is that the set of bacterial proteins that influence disease outcomes are enriched for secreted proteins that mediate interactions between pathogen and host macrophages. To test this hypothesis, we will collaborate with Cores A and B to use an integrative approach to combine genetic data from the M. tuberculosis GWAS datasets with genetic and proteomic screen to identify causal genes that mediate interactions with macrophages. We will use these same technologies to collaborate with Projects 2 and 3 to identify proteins/pathways responsible for host resistance and bacterial dissemination.

结核病（TB）是一种多方面疾病，在临床表现目的地有很大差异 带有单个病原体，结核分枝杆菌感染的产物。广泛的遗传多样性 负责临床结果差异的人类和结核分枝杆菌基因组代表 宿主 - 病原体相互作用的修饰，这是发病机理的关键。了解这些基础 异源反应将发现病毒和抗性的新机制，并会影响治疗 和诊断。不幸的是，研究感染差异结果的机制的挑战 在人类中，不仅包括识别宿主/病原体基因型与表型之间的相关性 人类人群，以及随后鉴定的疾病因果机制 需要在没有病原体的天然宿主生物的情况下在实验室中研究它们。这个程序需要 确定人类和人类的独特，持续基因组关联研究（GWAS）的优势 与两个不同人群中异质临床反应相关的病原体变异。 为了确定这些变化的机制，我们将采用一组强大的实验 测定，包括基于蛋白质组学的新扫描平台，以探测实验期间宿主反应 巨噬细胞感染与传统mRNA分析是正交的，以广泛寻找变化 与与这些临床菌株相关的疾病结局相关的宿主先天免疫病。 根据我们的初步数据，我们假设其中许多相互作用发生在感染期间的早期和 由结核分枝杆菌分泌的蛋白质介导。在此提案中，我们主要关注相关性 在两组独特的临床细菌之间，与之相关的菌株更大的传播 肺结核疾病和菌株更容易传播到体内的散落部位。一个 这两种菌株的意外主题是ESX分泌的结核病蛋白的普遍性 在结核分枝杆菌中表达的系统。结核分枝杆菌的ESX-1和ESX-5分泌系统都是 关键病毒确定细胞内生长和引起不同先天免疫反应所需的必需 在巨噬细胞感染期间。一个中心假设是影响疾病的细菌蛋白 结局丰富了介导病原体与宿主之间相互作用的分泌蛋白 巨噬细胞。为了检验这一假设，我们将与核心A和B合作使用综合方法 将结核分枝杆菌GWAS数据集的遗传数据与遗传和蛋白质组学结合 介导与巨噬细胞相互作用的因果基因。我们将使用这些相同的技术进行协作 通过项目2和3，可以识别负责宿主阻力和细菌的蛋白质/途径 传播。